Animal By-Products Order (Northern Ireland) 2002

Articles 7 and 8

SCHEDULE 1 Requirements for rendering plants

General requirements

1.  The approved premises shall be adequately separated from other buildings and from the public highway, except that they may be in the same building as a slaughterhouse provided that they are in a completely separate part.

2.  Animals and unauthorised persons shall not be permitted to enter the premises. Animals shall not be permitted to have access to unrendered animal by-products or any liquid from them. Preventive measures against birds, rodents, insects and other vermin shall be taken systematically.

3.  Floors shall be impervious, cleanable and be laid so that liquids drain away, and cannot seep from the unclean area into the clean area.

4.  The premises shall be drained by a waste water disposal system. Waste water originating in the unclean area shall be treated to ensure that no pathogens remain.

5.  Adequate lavatories, changing areas and washbasins shall be available for staff.

Clean and unclean areas

6.  There shall be a clean area and an unclean area, adequately separated. The unclean area shall be easy to clean and disinfect. There shall be a covered place (the reception area) to receive and store the unrendered animal by-products.

7.  Unrendered animal by-products shall be unloaded in the reception area and either—

(a) rendered immediately, or

(b) stored in the reception area and rendered without undue delay.

8.  If carcases are de-skinned or de-haired, there shall be adequate facilities in the unclean area for doing this, and there shall be a storage room for hides.

9.  Rendered material shall be handled, processed and stored in the clean area in such a way as to prevent recontamination. Rendered material shall not be allowed to come into contact with any unrendered animal by-products.

10.  Persons who have been in the unclean area shall not enter the clean area without first disinfecting or changing their footwear and changing their outer clothing. Equipment and utensils which have been in the unclean area shall not be taken into the clean area unless they have been suitably cleansed and disinfected.

Cleansing and disinfection facilities

11.  There shall be adequate facilities (including a water supply) provided to enable containers and vehicles (including their wheels) to be cleansed and disinfected in accordance with this paragraph. Vehicles (including their wheels) used for the transport of animal by-products shall be cleansed and disinfected before entering the clean area or (if they do not enter the clean area) before they leave the premises. Containers used for animal by-products shall be cleansed and disinfected after each use.

Equipment

12.  If the rendering method requires animal by-products to be reduced in size before rendering, the unclean area shall contain equipment to do this, and also equipment for loading the resultant material into the rendering unit.

13.  The premises shall have equipment capable of producing sufficient hot water and steam if these are used to render animal by-products.

14.—(1) Rendering premises shall be equipped with suitable rendering equipment. Where heat treatment is required, this installation shall be equipped with:

(a) measuring equipment to check temperature and, if necessary, pressure at critical points;

(b) recording devices to record continuously the results of measurements; and

(c) an adequate safety system to prevent insufficient heating.

15.  Installations and equipment shall be kept in a good state of repair. All measuring equipment shall be calibrated at regular intervals. A record shall be kept showing when re-calibrations were carried out.

Laboratories

16.  Where microbiological tests are required under Article 10, the premises shall either have their own laboratory authorised under this Order or make use of the services of a laboratory authorised under this Order.

Articles 2, 7, 8 and 17

SCHEDULE 2 Rendering

Part I Rendering Standards

Mammalian animal by-products

1.—(1) An operator shall render mammalian animal by-products in accordance with either—

(a) Method 1 of Part II of this Schedule; or

(b) any of the other methods in Part II of this Schedule if, after rendering, the resulting proteinaceous material is disposed of by burial, incineration, or a similar method which ensures that it will not enter any food or feed chain and will not be used as fertiliser.

(2) This paragraph shall not apply in relation to the rendering of the following mammalian material—

(a) low risk material for the production of petfood;

(b) hides, skins, hooves, horns or hair;

(c) blood or blood products;

(d) milk or milk products; or

(e) glands, tissues or organs for pharmaceutical use.

High risk material

2.  An operator shall render non-mammalian high risk material and mammalian high risk material specified in paragraph 1(2) in accordance with either—

(a) Method 1 of Part II of this Schedule; or

(b) any of the other methods in Part II of this Schedule (either in accordance with the parameters for the selected method set out in the Schedule or in accordance with the different parameters set out in the approval issued under Article 7) provided that the rendered material complies with the microbiological standards in Article 2(2).

Low-risk material

3.  An operator shall render non-mammalian low-risk material and mammalian low risk material specified in paragraph 1(2) in accordance with either—

(a) paragraph 2; or

(b) a method and parameters specified in the approval which ensure that the rendered material complies with the microbiological standards in Article 2(2).

Part-rendering

4.  Animal by-products may be part-rendered by any method approved by the Department if, after part-rendering, the material is disposed of in accordance with Article 5.

Gelatine and rendered fats

5.  The preceding methods and parameters shall not apply in relation to the rendering of animal by-products for the production of gelatine or rendered fats. Any animal by-products other than gelatine or rendered fats remaining after production shall be disposed of in accordance with Article 5.

Hides

6.  Hides shall be either rendered in accordance with the preceding provisions of this Schedule or salted using sodium chloride.

Re-rendering material

7.  If the required parameters are not achieved during any rendering operation, the material shall be rendered again so that those parameters are achieved.

Part II Rendering Methods

Method 1 Continuous or Batch Pressure

Reduction

1.  If the particle size of the animal by-products to be rendered is more than 50 mm, the animal by-products shall be reduced in size using equipment specified in the approval, set so that the particle size after reduction is no greater than 50 mm or such smaller size as the approval shall specify. The effectiveness of the equipment shall be checked daily and its condition recorded. If checks disclose the existence of particles larger than is permitted in the approval, the process shall be stopped and repairs made before the process is resumed.

Time and temperature

2.  After reduction the animal by-products shall be heated to a core temperature of more than 133°C for at least 20 minutes without interruption at a pressure of at least 3 bar.

3.  The rendering may be carried out in batch or continuous systems.

Method 2 Natural Fat Batch

Reduction

1.  If the particle size of the animal by-products to be rendered is more than 150 mm, the animal by-products shall be reduced in size using equipment specified in the approval, set so that the particle size after reduction is no greater than 150 mm or such smaller size as the approval shall specify. The effectiveness of the equipment shall be checked daily and its condition recorded. If checks disclose the existence of particles larger than is permitted in the approval, the process shall be stopped and repairs made before the process is resumed.

Time and temperature

2.  After reduction the animal by-products shall be heated to a core temperature greater than 100°C for at least 125 minutes, a core temperature greater than 110°C for at least 120 minutes and a core temperature greater than 120°C for at least 50 minutes.

3.  The rendering shall be carried out in a batch system.

4.  The animal by-products may be cooked such that the time-temperature requirements are achieved at the same time.

Method 3 Natural Fat

Continuous or Batch

Reduction

1.  If the particle size of the animal by-products to be rendered is more than 30 mm, the animal by-products shall be reduced in size using equipment specified in the approval, set so that the particle size after reduction is no greater than 30 mm or such smaller size as the approval shall specify. The effectiveness of the equipment shall be checked daily and its condition recorded. If checks disclose the existence of particles larger than is permitted in the approval, the process shall be stopped and repairs made before the process is resumed.

Time and temperature

2.  After reduction, the animal by-products shall be heated to a core temperature greater than 100°C for at least 95 minutes, a core temperature greater than 110°C for at least 55 minutes and a core temperature greater than 120°C for at least 13 minutes.

3.  The rendering may be carried out in batch or continuous systems.

4.  The animal by-products may be cooked such that the time-temperature requirements are achieved at the same time.

Method 4 Added Fat

Continuous or Batch

Reduction

1.  If the particle size of the animal by-products to be rendered is more than 30 mm, the animal by-products shall be reduced in size using equipment specified in the approval, set so that the particle size after reduction is no greater than 30 mm or such smaller size as the approval shall specify. The effectiveness of the equipment shall be checked daily and its condition recorded. If checks disclose the existence of particles larger than is permitted in the approval, the process shall be stopped and repairs made before the process is resumed.

Time and temperature

2.  After reduction the animal by-products shall be placed in a vessel with added fat and heated to a core temperature greater than 100°C for at least 16 minutes, a core temperature greater than 110°C for at least 13 minutes, a core temperature greater than 120°C for at least 8 minutes and a core temperature greater than 130°C for at least 3 minutes.

3.  The rendering may be carried out in batch or continuous systems.

4.  The animal by-products may be cooked such that the time-temperature requirements are achieved at the same time.

Method 5 Defatted

Continuous or Batch

Reduction

1.  If the particle size of the animal by-products to be rendered is more than 20 mm, the animal by-products shall be reduced in size using equipment specified in the approval set so that the particle size after reduction is no greater that 20 mm or such smaller size as the approval shall specify. The effectiveness of the equipment shall be checked daily and its condition recorded. If checks disclose the existence of particles larger than is permitted in the approval, the process shall be stopped and repairs made before the process is resumed.

Time and temperature

2.  After reduction the animal by-products shall be heated until they coagulate and then pressed so that fat and water are removed from the proteinaceous material. The proteinaceous material shall then be heated to a core temperature greater than 80°C for at least 120 minutes and a core temperature greater than 100°C for at least 60 minutes.

3.  The rendering may be carried out in batch or continuous systems.

4.  The animal by-products may be cooked such that the time-temperature requirements are achieved at the same time.

Method 6 Aquatic Animals

Combined Acidification and Heat Treatment

1.  The animal by-products shall be reduced to a size specified in the approval. They shall then be mixed with formic acid to reduce the pH to a level specified in the approval. They shall then be stored for a period specified in the approval.

2.  They shall then be heated to the temperature and time criteria specified in the approval.

3.  After heat treatment, the product shall be separated into liquid, fat and greaves by mechanical means. In order to obtain an animal protein concentrate, the liquid shall be pumped into two heat exchangers which are steam heated and equipped with vacuum chambers in order for its moisture to be removed in the form of water vapour. The greaves shall then be added to the protein concentrate.

Articles 2, 9 and 10

SCHEDULE 3 Sampling and Testing Methods

Part I Manner of Sampling

Method 1

1.  In accordance with the following table, samples of approximately equal size shall be extracted evenly from the whole of the rendered material. These samples shall then be divided into groups of approximately equal numbers, the number of groups being the number of aggregate samples specified in the table. The samples in each group shall then be mixed together to form aggregate samples.

Total quantity of rendered material consigned from the premises	Number of samples extracted	Number of aggregate samples obtained by mixing the relevant number of samples
Loose animal protein
up to 1 tonne	7	1
1–2.5 tonnes	7	2
2.5–10 tonnes	√20 × weight of sampled portion in tonnes	2
10–40 tonnes	√20 × weight of sampled portion in tonnes	3
over 40 tonnes	√20 × weight of sampled portion in tonnes	4
	(maximum – 40 incremental samples)
Bagged animal protein
1–16 bags	4	1
17–200 bags	√No. of bags of sampled portion	2
201–800 bags	√No. of bags of sampled portion	3
over 800 bags	√No. of bags of sampled portion	4
	(maximum – 40 incremental samples)

2.  Each aggregate sample shall be placed into a separate sterile receptacle and each shall be thoroughly mixed by stirring or shaking.

3.  Approximately equal amounts shall be taken from each aggregate sample and mixed so as to provide a single final sample of approximately 500 grammes. This final sample shall be transferred into a suitable sterile screw top container which shall then be sealed and marked to indicate its identity.

Method 2

1.  In accordance with the following table, samples of approximately equal size shall be extracted evenly from the whole of the rendered material. These samples shall then be divided into groups of approximately equal numbers, the number of groups being the number of aggregate samples specified in the table. The samples in each group shall then be mixed together to form aggregate samples.

Total consignments consigned from the premises	Number of samples extracted	Number of aggregate samples obtained by mixing the relevant number of samples
Loose or bagged animal protein
1–5 consignments	1 per consignment	1
6–10 consignments	1 per consignment	2
11–15 consignments	1 per consignment	3
Over 15 consignments	1 per consignment	4

For the purpose of this method “consignment” means the total quantity of rendered material loaded onto a single vehicle or trailer.

2.  Each aggregate sample shall be placed into a separate sterile receptacle and each shall be thoroughly mixed by stirring or shaking.

3.  Approximately equal amounts shall be taken from each aggregate sample and mixed so as to provide a single final sample of approximately 500 grammes. This final sample shall be transferred into a suitable sterile screw top container which shall then be sealed and marked to indicate its identity.

Part II Method for the Isolation of clostridium perfringens

Time of testing

1.  Tests shall be begun on receipt of the sample or on the first working day which allows this method to be completed. If the test is not begun on the day of receipt the sample shall be stored in a refrigerator at between 2°C and 8°C until required. If the sample has been refrigerated it shall be removed from the refrigerator and stored at room temperature for at least one hour before the test is started.

Samples

2.  Tests shall be carried out using two 10 gramme portions of each sample submitted for testing. Each 10 gramme sample shall be placed aseptically in a jar containing 90 ml Clostridium perfringens diluent consisting of 0.1% peptone and 0.8% sodium chloride at a pH of 7 and mixed thoroughly until the sample is evenly suspended.

Inoculations

3.  For each portion of the sample 1 ml of suspension/solution shall be transferred to a sterile 90 mm petri dish (in duplicate), to which 15 ml of egg yolk-free tryptose sulphite cycloserine agar (EY-free TSC agar)(1) at a temperature of 46°C ± 1°C shall be added and immediately gently mixed by swirling the dish with 5 clockwise and 5 anticlockwise circular movements.

4.  Once the agar has set, each agar plate shall be overlaid with a further 10 ml EY-free TSC agar at a temperature of 46°C ± 1°C. Once the overlay has set and with the plate lids uppermost the plates shall be incubated anaerobically at 37°C ± 1°C for 20 hours ± 2 hours.

Samples with colonies of Clostridium perfringens

5.  After incubation each set of duplicate plates shall be examined for colonies characteristic of Clostridium perfringens (black). The sample provisionally fails if any colonies characteristic of Clostridium perfringens are present, in which case the following procedure shall be followed to establish whether or not the colonies are Clostridium perfringens.

6.  In the case of each plate 10 characteristic colonies of Clostridium perfringens shall each be subcultured onto a further EY-free TSC agar plate. If there are less than 10 colonies on the plate, all characteristic colonies shall be subcultured onto further plates. The plates shall be incubated anaerobically at 37°C ± 1°C for 20 hours ± 2 hours.

7.  If the surface area of the plates is overgrown and it is not possible to select well isolated characteristic colonies, 10 suspect colonies shall each be subcultured onto duplicate EY-free TSC agar plates and incubated anaerobically at 37°C ± 1°C for 20 hours ± 2 hours.

8.  One characteristic colony from each plate shall be subcultured onto EY-free TSC agar and incubated anaerobically at 37°C ± 1°C for 20 hours ± 2 hours.

Subcultured colonies

9.  After incubation each plate shall be examined for colonies characteristic of Clostridium perfringens. All colonies characteristic of Clostridium perfringens shall be—

(a) stab inoculated into motility nitrate medium(2); and

(b) inoculated into either lactose gelatine medium(3) or charcoal gelatine discs(4) and incubated anaerobically at 37°C ± 1°C for 20 hours ± 2 hours.

Examination of subcultures

Motility

10.  The motility nitrate medium shall be examined for the type of growth along the stab line. If there is evidence of diffuse growth out into the medium away from the stab line, the bacteria shall be considered to be motile.

Reduction of nitrate to nitrite

11.  After examination of the motility nitrate medium 0.2ml to 0.5ml of nitrite detection reagent shall be added to it. The formation of a red colour confirms that the bacteria have reduced nitrate to nitrite. Cultures that show a faint reaction (i.e. a pink colour) should be discounted. If no red colour is formed within 15 minutes, a small amount of zinc dust shall be added and the plate allowed to stand for 15 minutes. If a red colour is formed after the addition of zinc dust no reduction of nitrate to nitrite has taken place.

Production of gas and acid from lactose and liquefaction of gelatine

12.  The lactose gelatine medium shall be examined for the presence of small gas bubbles in the medium.

13.  The lactose gelatine medium shall be examined for colour. A yellow colour indicates fermentation of lactose.

14.  The lactose gelatine medium shall be chilled for one hour at 2–8°C and then checked to see if the gelatine has liquefied. If the medium has solidified it shall be re-incubated anaerobically for a further 18–24 hours, the medium chilled for a further one hour at 2–8°C and again checked to see if the gelatine has liquefied.

15.  The presence of Clostridium perfringens shall be determined on the basis of the results from paragraphs 10 to 14. Bacteria which produce black colonies on EY-free TSC agar, are non-motile, reduce nitrate to nitrite, produce gas and acid from lactose and liquefy gelatine within 48 hours shall be considered to be Clostridium perfringens.

Control Tests

16.  Control tests shall be carried out each day that a test is initiated using

(a) Clostridium perfringens NCTC 10662(5) no more than seven days old at the time of use;

(b) Escherichia coli NCTC 10418 or equivalent not more than seven days old at the time of use; and

(c) rendered animal protein which is free of Clostridium perfringens.

17.  10 gramme portions of the rendered animal protein shall be placed aseptically in each of two jars containing 90 ml Buffered Peptone Water (BPW)(6) and mixed thoroughly until the samples are evenly suspended.

18.  One colony of Clostridium perfringens shall be placed in 10 ml BPW and mixed to form an even suspension. 0.1 ml of the suspension shall be added to the suspension in the preceding paragraph. This shall be repeated for Escherichia coli.

19.  These are then treated and examined in the same way as test samples. If no typical colonies are formed then that day’s testing shall be invalid and shall be repeated.

Part III Methods for the Isolation of Salmonella

A. Bacteriological Method

1.  Tests shall be begun on receipt of the sample or on the first working day which allows this method to be completed. If the test is not begun on the day of receipt the sample shall be stored in a refrigerator until required. If the sample has been refrigerated it shall be removed from the refrigerator and stored at room temperature for at least four hours before the test is started.

Day 1

2.  Tests shall be carried out in duplicate using two 25 gramme portions of each sample submitted for testing. Each 25 gramme sample shall be placed aseptically in a jar containing 225 ml Buffered Peptone Water (BPW) and incubated at 37°C ± 1°C for 18 hours.

Day 2

3.  0.1 ml from the jar of incubated BPW shall be inoculated into 10 ml Rappaport Vassiliadis (RV) broth(7) and incubated at 41.5°C ± 0.5°C for 24 hours.

Day 3

4.  The RV broth shall be plated out onto two 90 mm plates of Brilliant Green Agar(BGA)(8) or onto one 90mm plate of BGA and one 90 mm plate of Xylose Lysine Deoxycholate Agar (XLD)(9) using a 2.5 mm diameter loop. The plates shall be inoculated with a droplet taken from the edge of the surface of the fluid by drawing the loop over the whole of one plate in a zig zag pattern and continuing to the second plate without recharging the loop. The space between the loop streaks shall be 0.5 cm–1.0 cm. The plates shall be incubated at 37°C ± 1°C overnight.

5.  The residual RV broth shall be reincubated at 41.5°C ± 0.5°C for a further 24 hours.

Day 4

6.  The plates shall be examined and a minimum of 3 colonies from each plate showing suspicion of Salmonella growth shall be subcultured—

(a) onto a nutrient agar plate;

(b) onto a MacConkey agar plate(10); and

(c) into biochemical media suitable for the identification of Salmonella.

These media shall be incubated at 37°C ± 1°C overnight.

7.  The reincubated RV broth shall be plated out as described in paragraph 4.

Day 5

8.  The incubated composite media or equivalent shall be examined and the findings recorded, discarding cultures which are obviously not Salmonella. Slide serological tests shall be performed using Salmonella polyvalent “O” and polyvalent “H” (phase 1 and 2) agglutinating sera on selected suspect colonies collected from the nutrient agar or MacConkey agar plates. If reactions occur with one or both sera, the colonies shall be typed by slide serology and a subculture sent to one of the Department’s laboratories at either Food Science Division, Newforge Lane, Belfast, BT9 5PX or, Veterinary Science Division, Stoney Road, Belfast, BT4 3SD, for further typing.

9.  The plates referred to in paragraph 7 shall be examined and further action taken as in paragraphs 6 and 8.

B. Electrical conductance method

1.  Tests shall be begun on receipt of the samples of processed animal protein or on the first working day which allows the following method to be completed. If the test is not begun on the day of receipt the sample shall be stored in a refrigerator until required. If the sample has been refrigerated it shall be removed from the refrigerator and stored at room temperature for at least four hours before the test is started.

Day 1

2.  Tests shall be carried out in duplicate using two 25 gramme portions of each sample submitted for testing. Each 25 gramme sample shall be placed aseptically in a jar containing 225 ml Buffered Peptone Water (BPW) and incubated at 37°C ± 1°C for 18 hours.

Day 2

3.  The incubated BPW shall be added to Rappaport Vassiliadis Broth in electrical conductance cells or tubes to be inserted into electrical conductance cells. Detection of growth will utilise indirect impediometry as in the method of Donaghy and Madden (1993)(11). For cells or tubes containing more than 5 ml medium 0.2 ml of the BPW shall be added and for cells or tubes containing 5ml or less medium 0.1 ml of the BPW shall be added. Cells or tubes shall be connected to appropriate electrical conductance measuring equipment set to monitor and record changes in electrical conductance at 6 minute intervals over a 24 hour period. The temperature of cells and tubes shall be kept at 42°C.

Day 3

4.  At the end of the 24 hour period, the information recorded by the conductance measuring equipment shall be analysed and interpreted using criteria defined by the manufacturers of the equipment. Where a tube or cell is provisionally identified as being positive for Salmonella, the result shall be confirmed by subculturing the contents of the tube or cell onto two 90 mm plates of BGA or onto one 90 mm plate of BGA and one 90mm plate of Xylose Lysine Deoxycholate Agar (XLD) using a 2.5 mm diameter loop. The plates shall be inoculated with a droplet taken from the edge of the surface of the fluid by drawing the loop over the whole of one plate in a zig zag pattern and continuing to the second plate without recharging the loop. The space between the loop streaks shall be 0.5 cm–1.0 cm. The plates shall be incubated at 37°C ± 1°C overnight.

Day 4

5.  The plates shall be examined and a minimum of 3 colonies from each plate showing suspicion of Salmonella growth shall be subcultured—

(a) onto a nutrient agar plate;

(b) onto a MacConkey agar plate; and

(c) into biochemical media suitable for the identification of Salmonella.

These media shall be incubated at 37°C ± 1°C overnight.

Day 5

6.  The incubated composite media or equivalent shall be examined and the findings recorded, discarding cultures which are obviously not Salmonella. Slide serological tests shall be performed using Salmonella polyvalent “O” and polyvalent “H” (phase 1 and 2) agglutinating sera on selected suspect colonies collected from the nutrient agar or MacConkey plates. If reactions occur with one or both sera, the colonies shall be typed by slide serology and a subculture shall be sent to one of the Department’s laboratories at either Food Science Division, Newforge Lane, Belfast, BT9 5PX or Veterinary Science Division, Stoney Road, Belfast, BT4 3SD, for further typing.

Part IV Method for the isolation of enterobacteriaceae

1.  Tests shall be begun on receipt of the sample or on the first working day which allows this method to be completed. If the test is not begun on the day of receipt the sample shall be stored in a refrigerator until required at between 2°C and 8°C. If the sample has been refrigerated it shall be removed from the refrigerator and stored at room temperature for at least one hour before the test is started.

Samples

2.  Tests shall be carried out using five 10 gramme portions of each sample submitted for testing. Each 10 gramme sample shall be placed aseptically in a jar containing 90 ml Buffered Peptone Water (BPW) and mixed thoroughly until the sample is evenly suspended.

Inoculations

3.  For each portion of the sample 1 ml of solution shall be transferred to a sterile 90 mm petri dish (in duplicate). The plates shall be labelled to identify the portion of sample they were taken from. 15 ml of Violet Red Bile Glucose Agar (VRBGA)(12) at a temperature of 46°C ± 1°C shall be added to each petri dish and immediately gently mixed by swirling the dish with five clockwise and five anticlockwise circular movements.

4.  Once the agar has set, each agar plate shall be overlaid with a further 10 ml VRBGA at a temperature of 46°C ± 1°C. Once the overlay has set, the plates shall be inverted and incubated aerobically at 37°C ± 1°C for 20 hours ± 2 hours.

Samples with colonies of Enterobacteriaceae

5.  After incubation each set of duplicate plates shall be examined for colonies characteristic of Enterobacteriaceae (purple colonies 1–2mm in diameter). All characteristic colonies on each plate shall be counted and the arithmetic mean of the duplicate plates taken.

The sample provisionally fails if either—

(a) any arithmetic mean is above 30(13); or

(b) three or more arithmetic means are above 10;

in which case the following procedure shall be followed to establish whether or not the colonies are Enterobacteriaceae.

6.  After counting the colonies, characteristic colonies shall be taken at random from the agar plates, the number being at least the square root of the colonies counted. The colonies shall be subcultured onto a nutrient agar plate and incubated aerobically at 37°C ± 1°C for 20 hours ± 2 hours.

Examination of subcultures

7.  An oxidase test and a glucose fermentation test shall be performed on each of the five subcultured colonies. Colonies which are oxidase-negative and glucose fermentation-positive shall be considered to be Enterobacteriaceae.

8.  If not all of the colonies prove to be Enterobacteriaceae, the total count in paragraph 5 shall be reduced in proportion prior to establishing whether or not the sample should fail.

Controls

9.  Control tests shall be carried out each day that a test is initiated using—

(a) Escherichia coli NCTC 10418 no more than seven days old at time of use; and

(b) rendered animal protein which is free of Enterobacteriaceae.

10.  A 10 gramme portion of the rendered animal protein shall be placed aseptically in a jar containing 90 ml BPW and mixed thoroughly until the sample is evenly suspended.

11.  One colony of Escherichia coli shall be placed in 10 ml BPW and mixed to form an even suspension. 0.1 ml of the suspension shall be added to the suspension in the preceding paragraph.

12.  This is then treated and examined in the same way as test samples. If no typical colonies are formed then that day’s testing shall be invalid and shall be repeated.

Articles 17, 18 and 19

SCHEDULE 4 Requirements for knackers' yards

General requirements

1.  The approved premises must be adequately separated from the public highway and other premises such as slaughterhouses.

2.  Preventive measures against birds, rodents, insects and other vermin shall be taken systematically.

3.  Floors must be impervious, capable of being cleansed and disinfected and laid so that liquids drain into a waste-water disposal system and cannot seep from the unclean area to the clean area.

4.  Adequate lavatories, changing rooms and washbasins shall be available for staff.

Clean and unclean areas in premises producing feeding stuffs for animals whose flesh is not intended for human consumption

5.—(1) In premises producing feeding stuffs for animals whose flesh is not intended for human consumption there shall be a clean area and an unclean area, adequately separated. The unclean area shall be easy to clean and disinfect. It shall have a covered place (the reception area) to receive and store the untreated animal by-products.

(2) Untreated animal by-products shall be unloaded in the reception area and either—

(a) treated immediately; or

(b) stored in suitable containers in the reception area and treated without undue delay.

(3) Treated animal by-products shall be handled and stored in a clean area in such way as to prevent recontamination. Treated animal by-products shall not be allowed to come into contact with any untreated animal by-products.

Reception and storage facilities in premises not producing feeding stuffs

6.—(1) In premises not producing feeding stuffs for animals whose flesh is not intended for human consumption there shall be a covered place (the reception area) to receive and store the animal by-products. The premises shall be easy to clean and disinfect.

(2) Animal by-products shall be unloaded in the reception area and stored in suitable containers until disposal. They shall be disposed of without undue delay.

Hides

7.  If carcases are de-skinned or de-haired there shall be adequate facilities for doing this, and there shall be a storage room for hides. Hides shall be salted using sodium chloride.

Cleansing and disinfection facilities

8.—(1) There shall be adequate facilities (including a water supply) provided to enable containers and vehicles (including their wheels) to be cleansed and disinfected in accordance with this paragraph.

(2) Containers used for animal by-products shall be cleansed and disinfected after each use.

(3) In premises producing feeding stuffs for animals whose flesh is not intended for human consumption, vehicles (including their wheels) used for the transport of animal by-products shall be cleansed and disinfected before entering the clean area or (if they do not enter the clean area) before they leave the premises.

(4) In premises which do not produce feeding stuffs, vehicles (including their wheels) used for the transport of animal by-products shall be cleansed and disinfected before they leave the premises.

Repair of installations

9.  Installations and equipment shall be kept in a good state of repair and any measuring equipment shall be calibrated at regular intervals. A record shall be kept showing when re-calibration was carried out.

Products of knackers' yards

10.  A product of a knacker’s yard shall be either—

(a) disposed of without delay in accordance with Article 5; or

(b) treated in accordance with paragraph 11 and marketed locally within Northern Ireland for feeding animals whose flesh is not intended for human consumption.

Feeding stuffs

11.—(1) Animal by-products intended for use as feeding stuff for animals whose flesh is not intended for human consumption shall be either—

(a) sterilised by being boiled or steamed under pressure until every piece of meat is cooked throughout, or

(b) denatured by being stained with a solution of the colouring agent Black PN or Brilliant Black BN (E 151, Colour Index 197 No. 28440), the solution being of such a strength that the colouring on the stained meat is clearly visible and applied so that the whole surface of all pieces of meat have been covered with the solution either by immersing the meat in, or spraying or otherwise applying, the solution and, in the case of a piece of meat weighing 25 kilogrammes or more, applying the solution after the surface of the meat has been opened by multiple and deep incisions; or

(c) reduced to a particle size of no more than 50 mm and then heated to a core temperature of more than 133°C at a pressure of at least 3 bar for at least 20 minutes without interruption.

(2) Feeding stuffs shall be packaged before distribution and sale and the packaging shall include the name and address of the knacker’s yard and be clearly and legibly marked “Not for human consumption”.

(1)

EY- free TSC agar. See Hauschild, A.H.W. and Hilsheimer, R. (1974) Applied Microbiology 27:78–82. American Society for Microbiology, 1913 1 St N.W., Washington DC 20006, USA

(2)

Motility nitrate medium. See Hauschild AHW, Gilbert RJ, Harmon SM, O'Keefe MF, Vahlefeld R, (1997) ICMSF Methods Study VIII, Canadian Journal of Microbiology 23, 884–892. National Research Council of Canada, Ottawa ON K1A OR6, Canada

(3)

Lactose gelatine medium. See Hauschild AHW, Gilbert R J, Harmon S M, O'Keefe M F, Vahlefeld R, (1997) ICMSF Methods Study VIII, Canadian Journal of Microbiology 23, 884–892

(4)

Charcoal gelatine discs. See Mackie and McCartney, (1996) Practical Medical Microbiology 14,509. Churchill Livingstone, Robert Stevenson House, 1–3 Baxter’s Place, Leith Walk, Edinburgh EH1 3AF

(5)

The National Collection of Type Cultures, Central Public Health Laboratory, 61 Colindale Avenue, London NW9 5HT.

(6)

Buffered Peptone Water. See Edel, W. and Kampelmacher, E.H. (1973) Bulletin of World Health Organisation, 48: 167–174, World Health Organisation Distribution and Sales, CH-1211, Geneva 27, Switzerland (ISSN 0042-9686)

(7)

Rappaport Vassiliadis Broth. See Vassiliadis, P., Pateraki, E., Papaiconomou, N., Papadakis, J. A,. and Trichopoulos, D. (1976) Annales de Microbiologie (Institut Pasteur) 127B; 195–200. Elsevier, 23rue Linois, 75724 Paris, Cedex 15, France

(8)

Brilliant Green Agar. See Edel, W and Kampelmacher, E.H. (1969) Bulletin of World Health Organisation, 41:297–306, World Health Organisation Distribution and Sales, CH-1211, Geneva 27, Switzerland (ISSN 0042-9686)

(9)

Xylose Lysine Deoxycholate Agar. See Taylor, W.I. (1965) American Journal of Clinical Pathology, 44:471–475, Lippincott and Raven, 227 E. Washington Street Philadelphia PA19106, USA

(10)

MacConkey agar. See (1963) International Standards for Drinking Water. World Health Organisation Distribution and Sales, CH-1211, Geneva 27, Switzerland

(11)

Donaghy and Madden. See Donaghy, J.A. and Madden R.H. (1993) International Journal of Food Microbiology. 17;281–288

(12)

Violet Red Bile Glucose Agar-See Mossel, D.A.A., Eelderink, I. Koopmans, M., van Rossem, F. (1978) Laboratory Practice 27 No. 12 1049–1050; Emap Maclaren, PO Box 109, Maclaren House, 19 Scarbrook Road, Croydon CR9 1QH.

(13)

An arithmetic mean of 30 is equivalent to 3×10² colony forming units per gramme of original sample